An histochemical approach to characterization of anionic constituents in mast cell secretory granules

Ehud Skutelsky*, Tanya Shoichetman, Han Hammel

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

We used cationized colloidal gold in order to investigate the distribution of anionic sites in different secretory granules of rat and mouse mast cells. The localization of the anionic sites was performed by post-embedding labeling of thin sections of rat peritoneal cells or mouse skin tissue, fixed in Karnovsky's fixative and OsO4 and embedded in Araldite or LR white, respectively. In all cases anionic sites were demonstrated with a high density variation depending on cell type. In all mast cell secretory granules we have observed the highest density (ca. 500-900 gold particles/μm2), while in other peritoneal cell granules it was about 10 times less (ca. 40-80 gold particles/μm2). Pretreatment of the LR white sections with heparinase I and III resulted in a reduction of 97% and 72%, respectively, in the binding of the gold particles to the granules, indicating that the majority of the gold binding reactivity is due to heparin. Correlation of section profile area with labeling density revealed that the smaller granules were significantly more labeled when compared to the larger profiles. On the basis of these observations it seems that a post-translational change (mainly sulfation of heparin) of secretory content influences the granule anionic charge and thus may affect the intragranule buffer capacity.

Original languageEnglish
Pages (from-to)453-458
Number of pages6
JournalHistochemistry and Cell Biology
Volume104
Issue number6
DOIs
StatePublished - Dec 1995

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